Protein adsorption of Sephacryl S-type gels using high concentrations of ammonium sulfate

Abstract

A variety of proteins absorbed to Sephacryl gels, S-200, S-500 and S-1000 and Fractogel TSK-HW65 (F) in the presence of high concentrations of ammonium sulfate. The adsorption of these proteins to individual gel matrices showed some variations. The strength of adsorption of cytochrome c, myoglobin and chymotrypsinogen A to Sephacryl S-200 increased with temperature and pH, suggesting that hydrophobic interaction is involved in the adsorption. The chromatographic behaviour of these three proteins on Sephacryl S-type gels and Fractogel varied with the ammonium sulfate concentration of the eluant buffer. At 30% saturation, cytochrome c and myoglobin eluted together and followed by chymotrypsinogen A. At 45% saturation, however cytochrome c eluted before myoglobin and chymotrypsinogen A remained tightly bound to the column. Temperature, pH, flow-rate and glycerol were some of the factors affecting the separation of these three proteins on Sephacryl S-200.